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Casein and rice protein hydrolysates: molecular weight distribution analysis and in vitro potential allergenicity assessment

Autor Puigjaner, J.; Recio, Isidra ; Belloque, Josefina
Fecha de publicación 2010
Citación ESPGHAN 2010
Resumen[Objectives and Study]: Infant Formulas for infants with cow's milk protein allergy (CMA) are be based on extensively hydrolysed milk protein or hydrolysed rice protein. The goal of this study was to characterize molecular weight distribution of two casein and two rice hydrolysates and its IgE reactivity [Methods]: Molecular size distribution was measured with HPLC Size exclusion chromatography (SEC) (GE Healthcare, Superdex 30 pg Column) with Phosphate Buffer 50mM NaCl 0,15 M, pH = 7,2, Flux 1 ml/min), and detection at 214 nm. Absolute molecular weight was determined by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry method (MALDI-TOF), a rapid and sensitive quantitative method with high resolution for peptides. The IgE reactivity was evaluated using sera of patients with clinically demonstrated allergy to cows milk proteins that contains at least 20 KU/L of specific IgE antibodies towards milk proteins, measured using FEIACAP System (Pharmacia diagnostics, Uppsala, Sweden). IgE binding was measured using an indirect ELISA coupled to a signal amplification system. [Results]: SEC and MALDI-TOF allowed us to characterize hydrolysates in terms of peptide size abundance distribution. The percentage of peptides with a molecular weight under 2.000 Daltons (Da) varied from 85% to 95% Absolute maximum molecular weights detected by MALDI-TOF ranged from 1.968 to 2.685 Da. Hydrolysates presented no reactivity against IgE in serum of allergic patients when they were used at concentrations similar to native protein (0.0025 mg/ml). However, when they were used in concentrated form (1000x) they showed different degrees of reactivity. [Conclusion]: Precise molecular weight characterization and IgE in vitro reactivity may be useful tools to evaluate the appropriateness of protein hydrolysates for formula feeding of infants with cow's milk protein allergy (CMA) as screening test previous to clinical trials in humans.
Descripción Resumen del póster presentado al 43rd Annual Meeting of European Society for Paediatric Gastroenterology, Hepatology and Nutrition celebrado en Estambul (Turquia) del 9 al 12 de junio de 2010.-- et al.
URI http://hdl.handle.net/10261/113960
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